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Fig. 2 | Journal of Biomedical Science

Fig. 2

From: Long-term expansion of directly reprogrammed keratinocyte-like cells and in vitro reconstitution of human skin

Fig. 2

Selection, characterization, and expansion of BN-iKCs. a Schematic diagram of the selection of ITGA6+KRT15+ iKCs. b Immunostaining of selected ITGA6+KRT15+ BN-iKCs (BN28–2, BN28–5, and BN28–6) at passage 3 with specific antibodies against ITGA6 and KRT15. Nuclei were counterstained with DAPI. Images on the right are magnified views of the boxed areas in images on the left. Scale bars = 200 μm. c RT-PCR analysis of stem cell marker expression in pKCs, urine cells, and BN-iKCs. d Doubling time of BN-iKCs and BNK21–11 cells during long-term expansion. Cells were seeded at a density of 5 × 104 cells per well in a 6-well plate. Data represent the mean ± SEM. * P < 0.05. e Colony-forming assay of BN-iKCs during long-term expansion. BN-iKCs were seeded at a density of 0.25 × 104 cells per well in a 6-well plate with a 3 T3-J2 feeder layer and cultured for 1 week. The plates were stained with crystal violet

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